N-乙酰半胱氨酸对山羊子宫内膜基质细胞增殖及相关基因表达的影响

doi:10.11843/j.issn.0366-6964.2022.01.014

摘要/Abstract

摘要： 旨在探究N-乙酰半胱氨酸（N-acetyl-L-cysteine，NAC）对山羊子宫内膜基质细胞（goat endometrial stromal cells，gESCs）的影响。本研究以山羊子宫内膜基质细胞为对象，体外培养基中添加浓度梯度分别为100、200、400 μmol·L^-1的NAC，将0 μmol·L^-1 NAC视为空白组，运用CCK-8法筛选NAC的最适浓度并判断其对细胞的增殖情况，随后通过DCFH-DA探针、RT-qPCR分别检测最适浓度NAC对细胞内活性氧含量、细胞周期、增殖相关因子及山羊繁殖性能相关基因TGF-β1、TGF-β3 mRNA表达水平的影响。结果表明：200 μmol·L^-1 NAC浓度较其它梯度对山羊子宫内膜基质细胞增殖的影响更为显著，此外，200 μmol·L^-1 NAC较空白组降低了细胞ROS的含量（P<0.01），从而延缓细胞的衰老。RT-qPCR结果显示：200 μmol·L^-1 NAC添加能极显著提高细胞周期及增殖相关因子CyclinA1、CyclinE、PCNA mRNA的表达水平（P<0.01），降低细胞周期因子CyclinD2的mRNA表达量（P<0.01）；并极显著降低转化生长因子家族TGF-β1基因（P<0.01）、显著降低TGF-β3基因（P<0.05）的mRNA表达水平。故本研究推测200 μmol·L^-1 NAC可能通过调控细胞中CyclinA1、CyclinE、PCNA、CyclinD2的表达水平来促进细胞的增殖并通过调控转化生长家族因子TGF-β1、TGF-β3的表达水平影响山羊的繁殖性状，为进一步研究NAC对山羊繁殖性状的影响机理提供理论基础。

关键词: N-乙酰半胱氨酸, gESCs, 细胞增殖, 基因表达, 繁殖性能

Abstract: The aim was to explore the effect of N-acetyl-L-cysteine (NAC) on goat endometrial stromal cells (gESCs). In this study, goat endometrial stromal cells were used as the object. Three concentration gradients of 100,200,400 μmol·L^-1 NAC were added to the culture medium. The 0 μmol·L^-1 NAC was regarded as the blank group and CCK -8 method was used to screen the optimal concentration of NAC and judge its effect on cell proliferation.Then DCFH-DA probe and RT-qPCR were used to detect the optimal concentration of NAC on the intracellular reactive oxygen content, cell cycle and proliferation related factors, and goat reproductive performance-related genes TGF-β1 and TGF-β3 mRNA expression levels. The results showed that the concentration of 200 μmol·L^-1 NAC had a more significant effect on the proliferation of goat endometrial stromal cells than other gradients. In addition, 200 μmol·L^-1 NAC reduced the ROS content of cells compared with the blank group(P<0.01), thereby delaying cell senescence. RT-qPCR results showed that the addition of 200 μmol·L^-1 NAC significantly increased the expression levels of cell cycle and proliferation-related factors CyclinA1, CyclinE, PCNA mRNA (P<0.01), and reduced the mRNA expression of cell cycle factor CyclinD2(P<0.01); and extremely significantly reduced the TGF-β1 gene(P<0.01) of the transforming growth factor family and significantly reduced the mRNA expression level of the TGF-β3 gene(P<0.05). Therefore, this study speculates that 200 μmol·L^-1 NAC may promote cell proliferation by regulating the expression levels of CyclinA1, CyclinE, PCNA, and CyclinD2 in cells, and affect goat reproduction traits by regulating the expression levels of transforming growth family factors TGF-β1 and TGF-β3. This study provide a theoretical basis for further research on the mechanism of NAC’s influence on goat reproductive traits.

Key words: N-acetylcysteine, gESCs, cell proliferation, gene expression, reproductive performance

中图分类号:

S827.3

杨沛方, 陈祥, 周志楠, 唐文, 张艳, 惠茂茂. N-乙酰半胱氨酸对山羊子宫内膜基质细胞增殖及相关基因表达的影响[J]. 畜牧兽医学报, 2022, 53(1): 141-150.

YANG Peifang, CHEN Xiang, ZHOU Zhinan, TANG Wen, ZHANG Yan, HUI Maomao. The Effect of N-acetylcysteine on Proliferation and Related Gene Expression of Goat Endometrial Stromal Cells in vitro[J]. Acta Veterinaria et Zootechnica Sinica, 2022, 53(1): 141-150.

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